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5. Recurring infection by crayfish plague pathogen only marginally affects survival and growth of marbled crayfish

Author

Dobrović, Ana, primary, Geček, Sunčana, additional, Klanjšček, Tin, additional, Haberle, Ines, additional, Dragičević, Paula, additional, Pavić, Dora, additional, Petelinec, Ana, additional, Boštjančić, Ljudevit Luka, additional, Bonassin, Lena, additional, Theissinger, Kathrin, additional, and Hudina, Sandra, additional

Published
2022
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6. Nezaustavljiv: kronična infekcija marginalno utječe na preživljavanje i rast invazivne strane vrste mramornog raka

Author

Hudina, Sandra, Dobrović, Ana, Geček, Sunčana, Klanjšček, Tin, Haberle, Ines, Dragičević, Paula, Pavić, Dora, Petelinec, Ana, Boštjančić, Ljudevit Luka, Bonassin, Lena, Theissinger, Kathrin, Caput Mihalić, Katarina, Mičetić Stanković, Vlatka, Urlić, Inga, Mešić, Armin, and Kružić, Petar

Subjects
invazivne strane vrste, troškovi imunosnog odgovora, infekcija, limitacija hrane
Abstract

Invazivne strane vrste predstavljaju prijetnju raznolikosti slatkovodnih ekosustava i populacijama zavičajnih vrsta slatkovodnih rakova, primarno zbog prijenosa bolesti račje kuge. No, kronična infekcija račjom kugom može i kod tolerantne invazivne strane vrste dovesti do kompromisa u raspodijeli energije između ulaganja u imunosni odgovor i rasta i reprodukcije, što u kombinaciji s drugim nepovoljnim okolišnim uvjetima može utjecati na njezin invazivni uspjeh. U ovom smo istraživanju ispitali da li imunosni odgovor na infekciju patogenom račje kuge negativno utječe na rast i preživljavanje juvenilnih jedinki invazivne strane vrste mramornog raka (Procambarus virginalis). U sklopu eksperimentalnih istraživanja, jedinke su izložene različitim koncentracijama patogena (2) i različitim režimima hranjenja (2). U svim pokusima, ponavljajuća infekcija rezultirala je smanjenim prirastom jedinki. Značajan porast mortaliteta inficiranih jedinki zabilježen je tek pri uvjetima značajne limitacije hrane, u grupi koja je bila vrlo blizu izgladnjivanja. S obzirom da su infekcije bile česte, a koncentracije patogena visoke, rezultati pokazuju kako je mramorni rak otporan na patogen Aphanomyces astaci i kako dodatni stresori (izgladnjivanje) marginalno povećavaju mortalitet inficiranih jedinki. Stoga, kronična infekcija račjom kugom vjerojatno neće imati značajan utjecaj na invazivni uspjeh mramornog raka, čak niti u nepovoljnim uvjetima.

Published
2022

7. Genetic diversity and structure of the noble crayfish populations in the Balkan Peninsula revealed by mitochondrial and microsatellite DNA markers

Author

Gross, Riho, primary, Lovrenčić, Leona, additional, Jelić, Mišel, additional, Grandjean, Frederic, additional, Ðuretanović, Simona, additional, Simić, Vladica, additional, Burimski, Oksana, additional, Bonassin, Lena, additional, Groza, Marius-Ioan, additional, and Maguire, Ivana, additional

Published
2021
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8. Genetic diversity of the stone crayfish Austropotamobius torrentium (Schrank, 1803) populations in Croatia

Author

Bonassin, Lena and Maguire, Ivana

Subjects
mikrosatelitni lokusi, PRIRODNE ZNANOSTI. Biologija, populacijska struktura, conservation, konzervacija, population structure, NATURAL SCIENCES. Biology, microsatellite loci
Abstract

Potočni rak, Austropotamobius torrentium (Schrank, 1803), je nativna europska slatkovodna vrsta iz porodice Astacidae koja nastanjuje manje vodotoke na višim nadmorskim visinama. Rakovi porodice Astacidae su ključne vrste slatkovodnih ekosustava, a brojnost njihovih populacija, uključujući i populacija potočnog raka, je pod utjecajem raznih čimbenika koji dovode do značajnog smanjenja broja i veličina populacija. U ovom istraživanju razvijeni su novi mikrosatelitni markeri specifični za potočnog raka te su upotrebljeni u istraživanju genetske raznolikosti populacija ove vrste na području Hrvatske. Od 35 novorazvijenih markera, nakon testiranja, njih četiri su potvrđeni kao prikladni te su, uz četiri prethodno razvijena, korišteni za istraživanje populacijske strukture. Istraživanje je provedeno na 437 jedinki iz 17 populacija korištenjem osam mikrosatelitnih lokusa. Rezultati pokazuju veliku raznolikost populacija koje su grupirane prema geografskoj poziciji i filogrupama određenih na temelju mitohondrijske DNA. Rezultati ovog rada pridonose novim spoznajama o potočnom raku te su primjenjivi u izradi adekvatnih planova zaštite i upravljanja. The stone crayfish, Austropotamobius torrentium (Schrank, 1803), is a native European freshwater species of the family Astacidae that inhabits smaller waterbodies at higher altitudes. Freshwater crayfish are keystone species of freshwater ecosystems. The abundance of their populations, including those of the stone crayfish, is influenced by various factors that lead to a significant decline in the number and size of populations. In this research, new microsatellite markers, specific for A. torrentium, were developed. After testing 35 newly developed markers, four were confirmed as suitable. In addition, four previously developed microsatellite markers were used to study population structure of A. torrentium in Croatia. The study was conducted on 437 individuals from 17 populations using eight microsatellite loci. The results show a large diversity of populations grouped according to their geographical position and to phylogroups determined based on mitochondrial DNA. The results of this research contribute to the knowledge on stone crayfish and are applicable in the development of adequate protection and management plans.

Published
2021

9. The Pontastacus leptodactylus (Astacidae) repeatome provides insight into genome evolution and reveals remarkable diversity of satellite DNA

Author

Boštjančić, Ljudevit Luka, Bonassin, Lena, Anušić, Lucija, Lovrenčić, Leona, Besendorfer, Višnja, Maguire, Ivana, Grandjean, Frederic, Austin, Christopher M, Greve, Carola, Hamadou, Alexander B, Mlinarec, Jelena, Boštjančić, Ljudevit Luka, Bonassin, Lena, Anušić, Lucija, Lovrenčić, Leona, Besendorfer, Višnja, Maguire, Ivana, Grandjean, Frederic, Austin, Christopher M, Greve, Carola, Hamadou, Alexander B, and Mlinarec, Jelena

Abstract

Pontastacus leptodactylus is a native European crayfish species found in both freshwater and brackish environments. It has commercial importance for fisheries and aquaculture industries. Up till now, most studies concerning P. leptodactylus have focused onto gaining knowledge about its phylogeny and population genetics. However, little is known about the chromosomal evolution and genome organization of this species. Therefore, we performed clustering analysis of a low coverage genomic dataset to identify and characterize repetitive DNA in the P. leptodactylus genome. In addition, the karyogram of P. leptodactylus (2n = 180) is presented here for the first time consisting of 75 metacentric, 14 submetacentric, and a submetacentric/metacentric heteromorphic chromosome pair. We determined the genome size to be at ~18.7 gigabase pairs. Repetitive DNA represents about 54.85% of the genome. Satellite DNA repeats are the most abundant type of repetitive DNA, making up to ~28% of the total amount of repetitive elements, followed by the Ty3/Gypsy retroelements (~15%). Our study established a surprisingly high diversity of satellite repeats in P. leptodactylus. The genome of P. leptodactylus is by far the most satellite-rich genome discovered to date with 258 satellite families described. Of the five mapped satellite DNA families on chromosomes, PlSAT3-411 co-localizes with the AT-rich DAPI positive probable (peri)centromeric heterochromatin on all chromosomes, while PlSAT14-79 co-localizes with the AT-rich DAPI positive (peri)centromeric heterochromatin on one chromosome and is also located subterminally and intercalary on some chromosomes. PlSAT1-21 is located intercalary in the vicinity of the (peri)centromeric heterochromatin on some chromosomes, while PlSAT6-70 and PlSAT7-134 are located intercalary on some P. leptodactylus chromosomes. The FISH results reveal amplification of interstitial telomeric repeats (ITRs) in P. leptodactylus. The prevalence of repetitive elements

Published
2021

10. Recurring infection by crayfish plague pathogen only marginally affects survival and growth of marbled crayfish.

Author

Dobrović, Ana, Geček, Sunčana, Klanjšček, Tin, Haberle, Ines, Dragičević, Paula, Pavić, Dora, Petelinec, Ana, Boštjančić, Ljudevit Luka, Bonassin, Lena, Theissinger, Kathrin, and Hudina, Sandra

Subjects
CRAYFISH, LIFE history theory, HAPLOGROUPS, INTRODUCED species, PATHOGENIC microorganisms, INFECTION
Abstract

Invasive alien crayfish threaten the diversity of freshwater ecosystems and native crayfish fauna. In Europe, this is largely due to transmission of the crayfish plague to susceptible native crayfish. Many invasive species tolerate crayfish plague, but the infection still has the potential to reduce the fitness of a tolerant host due to energy trade-offs between immune response maintenance and life-history traits, such as growth and reproduction. In combination with other unfavourable conditions, such a response could alter further invasion success of an otherwise successful crayfish invader. We examined whether repeated infection with one of the most virulent haplogroups of crayfish plague agent (Aphanomyces astaci) affects growth or survival of the juvenile marbled crayfish (Procambarus virginalis). Juveniles were exposed to i) two levels of pathogen concentrations, and ii) two different feeding regimes under the higher pathogen concentration. In all performed trials, repeated infection reduced growth rates, while the combination of recurring infection and food limitation significantly increased mortality. The average energy cost of the immune response was estimated at 12.07 J/day for individuals weighing 0.3 grams. Since infections were frequent and pathogen concentrations high, results suggest that marbled crayfish is resistant to A. astaci pathogen and its survival is only affected by adding the stress of food limitation. The survival of almost half of the individuals exposed to high pathogen loads and extreme food limitation indicates that chronic infection by crayfish plague is unlikely to be an important factor impeding invasion success of the marbled crayfish, even under harsh conditions. Our results add to the growing body of evidence that marbled crayfish has potential to become one of the most successful freshwater invaders. [ABSTRACT FROM AUTHOR]

Published
2022
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11. The Pontastacus leptodactylus (Astacidae) Repeatome Provides Insight Into Genome Evolution and Reveals Remarkable Diversity of Satellite DNA

Author

Boštjančić, Ljudevit Luka, primary, Bonassin, Lena, additional, Anušić, Lucija, additional, Lovrenčić, Leona, additional, Besendorfer, Višnja, additional, Maguire, Ivana, additional, Grandjean, Frederic, additional, Austin, Christopher M., additional, Greve, Carola, additional, Hamadou, Alexander Ben, additional, and Mlinarec, Jelena, additional

Published
2021
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12. Uloga kompatibilnih osmolita u odgovoru biljaka na osmotski stres

Author

Bonassin, Lena and Tkalec, Mirta

Subjects
PRIRODNE ZNANOSTI. Biologija, osmotski stres, NATURAL SCIENCES. Biology, abiotički stres, nedostatak vode, vodni potencijal, prolin, osmoliti
Abstract

Biljke su tijekom rasta i razvoja izložene različitim okolišnim uvjetima. Nepovoljni uvjeti mijenjaju fiziološke procese, metabolizam, rast i razvoj biljke te dovode do promjena na staničnoj i molekularnoj razini. Osmotski stres se javlja zbog abiotičkih faktora kao što su povišeni salinitet, suša ili ekstremne temperature. Tijekom takvih uvjeta narušen je vodni potencijal biljke. Kako bi se osmolarnost povećala i smanjio gubitak vode, dolazi do sinteze kompatibilnih osmolita koji su organske molekule niske molekulske mase. Biljke nakupljaju različite spojeve: aminokiseline, tercijarne sulfonijeve spojeve, kvarterne amonijeve spojeve, šećere i šećerne alkohole. Osmoliti se nakupljaju u citoplazmi, gdje održavaju osmotski potencijal i fiziološku aktivnost stanice te je njihova biosinteza pod utjecajem okolišnih signala. Takvi spojevi ne utječu na stanične reakcije, a pomažu u toleranciji stresnih uvjeta stabilizacijom makromolekula i staničnih struktura, detoksifikacijom reaktivnih oblika kisika i održavanjem redoks sustava u stanici. Genskom manipulacijom gena za sintezu osmolita moguće je stvoriti biljke koje akumuliraju osmolite u povišenim koncentracijama te tako poboljšati njihovu sposobnost tolerancije stresnih uvjeta. During growth and development plants are exposed to different environmental factors. Adverse conditions change the physiological processes, the metabolism, growth and development of the plant which affect the organism at a cellular and molecular level. Osmotic stress is a consequence of abiotic factors such as high salinity, drought or extreme temperatures. Those conditions disturb the water potential of the cell. To increase the potential and decrease water loss, compatible osmolytes are synthesized. Osmolytes are low molecular mass organic compounds. Plants accumulate different compounds: amino acids, tertiary sulphonium compounds, quaternary ammonium compounds, sugars and sugar alcohols which accumulate in the cytoplasm. Compatible osmolytes maintain the osmotic potential and physiological activity of the cell and are synthesized under control of environmental cues. Additionally, they stabilize the macromolecule and cell structure, scavenge reactive oxygen species and maintain the cell redox system without affecting the metabolic reactions of the cell. With genetic engineering of osmolytes biosynthetic pathways, it is possible to create plants accumulating osmolytes in high concentration which can improve their stress tolerance.

Published
2018

13. Filogeografska struktura ugroženog potočnog raka - nove spoznaje

Author

Jelić, Mišel, Boštjančić, Ljudevit Luka, Bonassin, Lena, Podnar, Martina, Slavevska- Stamenković, Valentina, Mirt, Martina, Maguire, Ivana, Kružić, Petar, Caput Mihalić, Katarina, Gottstein, Sanja, Pavoković, Dubravko, and Kučinić, Mladen

Subjects
Austropotamobius torrentium, jugoistočna Europa, konzervacija, bioraznolikost
Abstract

Potočni rak Austropotamobius torrentium (Schrank, 1803) jedan je od pet nativnih europskih vrsta rakova koje nastanjuju manje vodotokove na većim nadmorskim visinama središnje i jugoistočne Europe. U proteklim desetljećima uočeno je smanjenje populacija potočnih rakova uzrokovano antropogenskim pritiskom na njihova staništa, širenjem stranih vrsta rakova i klimatskim promjenama. Stoga se potočni rak smatra ugroženom vrstom, zaštićenom međunarodnim i nacionalnim zakonima. Poseban naglasak potrebno je staviti na razvoj konzervacijskih planova te upotpuniti saznanja o samoj vrsti, kako bi se razvila učinkovita zaštita. To uključuje i nedavna molekularno filogenetička istraživanja vrste A. torrentium koja su pokazala da unutar vrste postoji namanje 7 različitih monofiletskih haplogrupa s najvećom raznolikosti u sjevernim i središnjim Dinaridima. Dosad neistražene populacije potočnog raka iz Hrvatske, Slovenije i Makedonije uključene su u ovo istraživanje uz uzorke s više od 190 lokaliteta s ciljem da se pomoću mitohondrijskih (16S rRNA i COI) i jezgrinih (ITS2 regija) DNA biljega odrede filogenetski odnosi unutar vrste. Zabilježeno je ukupno 48 16S rRNA i 147 COI haplotipova te veći broj ITS2 alela. Filogenetska rekonstrukcija napravljena na sekvencama sva tri DNA biljega, pokazala je kongruentne topologije te nove haplogrupe nisu pronađene. Raspravljena je primjena dobivenih rezultata u budućim planovima očuvanja vrste A. torrentium. (The stone crayfish Austropotamobius torrentium (Schrank, 1803) is one of five native European crayfish species inhabiting smaller waterbodies at higher altitudes of central and south- eastern Europe. In the last decades severe declines in numbers of stone crayfish populations have been recorded, due to anthropogenic pressure on their habitats, spread of non-native crayfish species and climate change. Hence stone crayfish is considered threatened species, protected by international and national laws. The sound knowledge of species, including information on genetics, is a necessity to develop efficient strategies for its protection. Recent molecularphylogenetic research of A. torrentium revealed that within the species at least 7 distinct monophyletic haplogroups exist, with highest genetic diversity in the north-central Dinaric region. This research aimed to update previous results through the inclusion of previously unstudied populations of stone crayfish from Croatia, Slovenia and Macedonia. Samples from more than 190 localities were analysed either for mitochondrial (16S rRNA and COI) or nuclear (ITS2 region) DNA markers. A total of 48 16S rRNA and 147 COI haplotypes, and several ITS2 alleles were obtained. Phylogenetic relationships were reconstructed using all three datasets with different methods of phylogenetic inference showing congruent topologies and no new haplogroups. Applications of acquired results in the future A. torrentium conservation programs are discussed.)

Published
2018

14. Expression of C/EBP and Kr-h1 transcription factors under immune stimulation in the noble crayfish.

Author

Boštjančić, Ljudevit Luka, Dragičević, Paula, Bonassin, Lena, Francesconi, Caterina, Tarandek, Anita, Schardt, Leonie, Rutz, Christelle, Hudina, Sandra, Schwenk, Klaus, Lecompte, Odile, and Theissinger, Kathrin

Subjects
*GENETIC regulation, *GENE expression, *GENE regulatory networks, *ABDOMINAL muscles, *CRAYFISH, *PLANT cell walls
Abstract

[Display omitted] • β-1–3-glucan (laminarin) has a potential as an immunostimulant. • THC is increased following the immune stimulation with CBS and β-1–3-glucan. • Domain organisation of C/EBP and Kr-h1 is conserved in freshwater crayfish. • C/EBP and Kr-h1 are ubiquitously expressed in all tested tissues. • Immune stimulation with β-1–3-glucan leads to C/EBP up-regulation. Transcription factors (TFs) have an important role in the regulation of the gene expression network. The role of TFs in the immune response of freshwater crayfish is poorly understood, but leveraging the regulatory mechanisms of immune response could augment the resistance against the invasive oomycete pathogen, Aphanomyces astaci. Previous studies indicated that the TFs CCAAT/enhancer-binding protein (C/EBP) and putative Krüppel homolog-1 protein (Kr-h1) might play a role in immune and stress response of the noble crayfish (Astacus astacus). Here, we aimed to further characterise these two gene products to gain a better understanding of their evolutionary origin, domain organisation and expression patterns across different crayfish tissues. Furthermore, we conducted an immune stimulation experiment to observe the potential changes in the gene expression of C/EBP and Kr-h1 under immune challenge in different crayfish tissues. Our results showed that both C/EBP and Kr-h1 are closely related to other C/EBP s and Kr-h1 s in Malacostraca. Gene expression analysis revealed that both TFs are present in all analysed tissues, with higher expression of C/EBP in the gills and Kr-h1 in the abdominal muscle. Immune stimulation with laminarin (mimicking β-1–3-glucan in the oomycete cell wall) showed an activation of the crayfish immune system, with an overall increase in the total haemocyte count (THC) compared to untreated control and crayfish buffered saline (CBS) treatment. On the gene expression level, an up-regulation of the C/EBP gene was detected in the laminarin treated group in hepatopancreas and heart, while no changes were observed for the Kr-h1 gene. Our results indicate an early change in C/EBP expression in multiple tissues during immune stimulation and suggest its involvement in the immune response of the noble crayfish. [ABSTRACT FROM AUTHOR]

Published
2024
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15. Additional file 5 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 5: Results of species delimitation analyses performed on Austropotamobius torrentium COI dataset applying different methods (ABGD (lumper, splitter), TCS, bPTP, mPTP and GMYC).

16. Additional file 9 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 9: Material and methods extended. The detailed information about gene amplification and sequencing, phylogenetic reconstruction, time of divergence estimates, species delimitation and validation, and analyses of meristic characteristics. Additional file also includes list of used references.

17. Additional file 9 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 9: Material and methods extended. The detailed information about gene amplification and sequencing, phylogenetic reconstruction, time of divergence estimates, species delimitation and validation, and analyses of meristic characteristics. Additional file also includes list of used references.

19. Additional file 5 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 5: Results of species delimitation analyses performed on Austropotamobius torrentium COI dataset applying different methods (ABGD (lumper, splitter), TCS, bPTP, mPTP and GMYC).

23. Additional file 4 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 4: Estimation of divergence times based on Austropotamobius torrentium mitochondrial data set using three molecular clock and four geological calibrations.

24. Additional file 8 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 8: Map of proposed evolutionary significant units (ESUs)/cryptic subspecies for Austropotamobius torrentium. The map depicted in figure was produced in ArcGIS 10.3 program package and finished in the program package FreeHand MXa by authors of this study.

25. Additional file 4 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 4: Estimation of divergence times based on Austropotamobius torrentium mitochondrial data set using three molecular clock and four geological calibrations.

27. Additional file 8 of New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications

Author

Lovrenčić, Leona, Bonassin, Lena, Boštjančić, Ljudevit Luka, Podnar, Martina, Mišel Jelić, Klobučar, Göran, Jaklič, Martina, Slavevska-Stamenković, Valentina, Hinić, Jelena, and Maguire, Ivana

Subjects
14. Life underwater
Abstract

Additional file 8: Map of proposed evolutionary significant units (ESUs)/cryptic subspecies for Austropotamobius torrentium. The map depicted in figure was produced in ArcGIS 10.3 program package and finished in the program package FreeHand MXa by authors of this study.

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